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Dual targeting to immune checkpoints has achieved a better therapeutic efficacy than single targeting due to synergistic extrication of tumour immunity. However, most dual targeting strategies are usually antibody dependent which facing drawbacks of antibodies, such as poor solid tumour penetration and unsatisfied affinity. To meet the challenges, we engineered a cell membrane displaying a fusion protein composed of SIRPa and PD-1 variants, the high-affinity consensus (HAC) of wild-type molecules, and with which prepared nanovesicles (NVs). Through disabling both SIRPa/CD47 and PD-1/PD-L1 signalling, HAC NVs significantly preserved the phagocytosis and antitumour effect ofmacrophages and T cells, respectively. In vivo study revealed that HAC NVs had better tumour penetration than monoclonal antibodies and higher binding affinity to CD47 and PD-L1 on tumour cells compared with the NVs expressing wild-type fusion protein. Exhilaratingly, dual-blockade of CD47 and PD-L1 with HAC NVs exhibited excellent therapeutic efficacy and biosafety. This study provided a novel biomaterial against tumoural immune escape and more importantly an attractive biomimetic technology of protein delivery formulti-targeting therapies. [ABSTRACT FROM AUTHOR]

Mitogen-activated protein kinases (MAPKs, also known as MPKs) regulate diverse cellular and physiological functions, and dual-specificity MAPK phosphatases (MKPs) modulate MAPK signalling through MAPK dephosphorylation and inactivation. Due to lacking of overall understanding for the regulatory networks between Chrysanthemum morifolium MKPs (CmMKPs) and C. morifolium MAPKs (CmMPKs), we systematically studied the interactions between four groups of CmMPKs and eight identified CmMKPs in chrysanthemum and found that the interaction between the specific CmMKP and the specific CmMPK differed from those in other plants. Furthermore, the expression of CmMKP1 and CmMKP1-LIKE1 showed opposite trends during the development of chrysanthemum flower buds under salt treatment and Alternaria alternata inoculation, but these genes could interact with the same CmMPKs, providing insight into the subfunctionalization of paralogues. Amino acid variations (M87V, T277P and V6L) in dual-specificity protein phosphatases (DsPTP1)-LIKE1/2/3 changed the interactions of these proteins with the four CmMPK groups in chrysanthemum, providing evidence for the de/neofunctionalization of paralogues in polyploids, suggesting that we can identify the key functional sites of proteins by studying polyploid paralogues. [ABSTRACT FROM AUTHOR]

Antimicrobial peptides are a promising resource for developing novel antibiotic and even anticancer drugs. Here, a 28-mer polypeptide, Ranatuerin-2PLx (R2PLx), was identified from lyophilised skin secretions. The chemically synthetic replicates exhibited moderate and broadspectrum antimicrobial effect against various microorganisms including methicillin-resistant Staphylococcus aureus (MRSA, minimal inhibitory concentration = 256 μM). In addition, R2PLx was found to inhibit the proliferation of several tumour cells, especially showing more potent effect on prostate cancer cell, PC-3. The early cell apoptosis was observed in 6 h by Annexin V-FITC/propidium iodide staining, as well as the activation of Caspase-3 at 5 μM peptide concentration. R2PLx may therefore be promising for developing new therapeutic approach for cancer treatment. Moreover, the artificial deficiency of conserved rana-box loop or net positive charge in C-terminal domain notably reduced the biological activities of the truncated and substituted isoforms, respectively, suggesting for maintaining their biological potency of ranatuerin family requires both cysteine-bridged segment and cationincity within the loop domain in C-terminus. [ABSTRACT FROM AUTHOR]

Endothelial leakiness or permeability directly determines the access of any bionanotechnology to the target tissue site. Currently, cancer nanotechnology relies on tumor-derived endothelial leakiness, which suffers from unreliability, inhomogeneity of leakiness, and uncontrollability. Nanomaterials by themselves are capable of inducing endothelial leakiness (NanoEL) without any tumor involvement by targeting the endothelial cell junctions; this NanoEL phenomenon not well understood. Here, we showed that the negatively charged Au nanoparticles (NPs) induce significantly higher NanoEL than positively charged nanoparticles. We hypothesized and showed that in both in vitro and in silico models that cell junction targeting arose for the negatively charged particles due to a succession of repulsive-sedimentary interactions between the negative particle and the negatively charged glycocalyx found on the cell membrane surface. On the contrary, NPs with positive charges are attracted stably by the negatively charged glycocalyx and remained in situ for long enough, which eventually are endocytosized into the cells as contrasted to localization toward the cell junction. There are implications to how nanoparticle charges could be tuned to induce (or avoid) endothelial leakiness by design. [ABSTRACT FROM AUTHOR]

Background: Enhanced recovery programs have become an important focus of perioperative management. A few studies have demonstrated the efficacy of an enhanced recovery after surgery (ERAS, which includes optimized pain control, restricted I.V. fluids, early initiation of postoperative oral feeding, and enforced mobilization) protocol in patients undergoing radical gastrectomy. We investigated the feasibility and safety of laparoscopic radical gastrectomy within ERAS programs.Methods: In this single-center prospective randomized controlled trial conducted between September 2013 and August 2014, 149 consecutive locally advanced gastric cancer patients (T2-4, any N, M0) diagnosed by the CT scanning were allocated to either ERAS group (N = 73) or conventional pathway group (N = 76). The same surgical technique was used in both groups, that is, laparoscopic gastrectomy with D2 lymphadenectomy and R0 resection. Intergroup differences were evaluated for clinical parameters and C-reactive protein for testing tissue injury.Results: ERAS combined with laparoscopic gastrectomy was observed in our study. Recovery parameters such as time to return to normal diet (days) 1.90 ± 0.71 versus 3.52 ± 0.81, P = .003 and time to the first defecation (days) 2.97 ± 1.23 versus 5.20 ± 1.81, P = .015 were measured. The post hospital stay (days) in ERAS and the conventional care group were 6.38 ± 2.04 and 8.62 ± 2.87, P < .001, respectively. No statistically significant intergroup differences were observed in terms of postoperative complications and C-reactive protein levels. One patient in the ERAS group was readmitted because of anastomotic leakage. No instances of deaths were reported during the 30-day follow-up period.Conclusion: This study demonstrates the feasibility and safety of ERAS protocol in advanced gastric cancer patients undergoing laparoscopic radical gastrectomy and was associated with shorter duration of hospital stay. ( Clinicaltrials.gov Identifier No NCT02348229). [ABSTRACT FROM AUTHOR]

Glioma is a type of tumor derived from glial cells, which is associated with a high level of incidence and mortality. At present, the generation of a fast and efficient method to evaluate the malignancy grade of glioma is required. Cancer stem cells (CSCs) are currently attracting attention in oncological studies; therefore, the present study aimed to investigate novel biomarkers of glioma CSCs, in order to provide new criteria for the grading of glioma. The mRNA expression levels of CD133, (sex determining region Y)-box 2, nestin, vascular endothelial growth factor (VEGF) and phosphoinositide-3-kinase (PI3K) were detected in 15 human samples of high-malignancy glioma and 12 human samples of low-malignancy glioma in vitro. The mRNA expression levels of VEGF and PI3K were higher in the high-malignancy group, as compared with in the low-malignancy group. In conclusion, the mRNA expression levels of VEGF and PI3K in glioma CSCs may be considered a novel criteria for the grading of glioma. [ABSTRACT FROM AUTHOR]

Bacillus subtilis as the Gram-positive model bacterium has been widely used in synthetic biology and biotechnology while the regulatory RNA tools for B. subtilis are still not fully explored. Here, a bottom-up approach is proposed for designing artificial trans-acting sRNAs. By engineering the intrinsic sRNA SR6, a minimized core scaffold structure consisting of an 8 bp stem, a 4 nt loop, and a 9 nt polyU tail was generated and proven to be sufficient for constructing sRNAs with strong repression activity (83%). Moreover, we demonstrate this artificial sRNA system functions well in an hfq-independent manner and also achieves strong repression efficiency in Escherichia coli (above 80%). A structure-based sRNA design principle was further developed for the automatic generation of custom sRNAs with this core scaffold but various sequences, which facilitates the manipulation and avoids structure disruption when fusing any base-pairing sequence. By applying these auto-designed sRNAs, we rapidly modified the cell morphology and biofilm formation, and regulated metabolic flux toward acetoin biosynthesis. This sRNA system with cross-species regulatory activities not only enriched the gene regulation toolkit in synthetic biology for B. subtilis and E. coli but also enhanced our understanding of trans-acting sRNAs. [ABSTRACT FROM AUTHOR]

Bacteria exhibit a rich repertoire of RNA molecules that intricately regulate gene expression at multiple hierarchical levels, including small RNAs (sRNAs), riboswitches, and antisense RNAs. Notably, the majority of these regulatory RNAs lack or have limited protein-coding capacity but play pivotal roles in orchestrating gene expression by modulating transcription, post-transcription or translation processes. Leveraging and redesigning these regulatory RNA elements have emerged as pivotal strategies in the domains of metabolic engineering and synthetic biology. While previous investigations predominantly focused on delineating the roles of regulatory RNA in Gram-negative bacterial models such as Escherichia coli and Salmonella enterica, this review aims to summarize the mechanisms and functionalities of endogenous regulatory RNAs inherent to typical Gram-positive bacteria, notably Bacillus subtilis. Furthermore, we explore the engineering and practical applications of these regulatory RNA elements in the arena of synthetic biology, employing B. subtilis as a foundational chassis. [ABSTRACT FROM AUTHOR]

Super-enhancers (SEs) are clusters of enhancers marked by extraordinarily high and broad chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) signals for H3K27ac or other transcription factors (TFs). SEs play pivotal roles in development and oncogenesis. Epstein-Barr virus (EBV) super-enhancers (ESEs) are co-occupied by all essential EBV oncogenes and EBV-activated NF-κB subunits. Perturbation of ESEs stops lymphoblastoid cell line (LCL) growth. To further characterize ESEs and identify proteins critical for ESE function, MYC ESEs were cloned upstream of a green fluorescent protein (GFP) reporter. Reporters driven by MYC ESEs 525 kb and 428 kb upstream of MYC (525ESE and 428ESE) had very high activities in LCLs but not in EBV-negative BJAB cells. EBNA2 activated MYC ESE-driven luciferase reporters. CRISPRi targeting 525ESE significantly decreased MYC expression. Genome-wide CRISPR screens identified factors essential for ESE activity. TBP-associated factor (TAF) family proteins, including TAF8, TAF11, and TAF3, were essential for the activity of the integrated 525ESE-driven reporter in LCLs. TAF8 and TAF11 knockout significantly decreased 525ESE activity and MYC transcription. MEF2C was also identified to be essential for 525ESE activity. Depletion of MEF2C decreased 525ESE reporter activity, MYC expression, and LCL growth. MEF2C cDNA resistant to CRIPSR cutting rescued MEF2C knockout and restored 525ESE reporter activity and MYC expression. MEF2C depletion decreased IRF4, EBNA2, and SPI1 binding to 525ESE in LCLs. MEF2C depletion also affected the expression of other ESE target genes, including the ETS1 and BCL2 genes. These data indicated that in addition to EBNA2, TAF family members and MEF2C are essential for ESE activity, MYC expression, and LCL growth. IMPORTANCE SEs play critical roles in cancer development. Since SEs assemble much bigger protein complexes on enhancers than typical enhancers (TEs), they are more sensitive than TEs to perturbations. Understanding the protein composition of SEs that are linked to key oncogenes may identify novel therapeutic targets. A genome-wide CRISPR screen specifically identified proteins essential for MYC ESE activity but not simian virus 40 (SV40) enhancer. These proteins not only were essential for the reporter activity but also were also important for MYC expression and LCL growth. Targeting these proteins may lead to new therapies for EBV-associated cancers. [ABSTRACT FROM AUTHOR]

Epstein-Barr virus (EBV) infection of human primary resting B lymphocytes (RBLs) leads to establishment of lymphoblastoid cell lines (LCLs) that can grow indefinitely in vitro. EBV transforms RBLs through expression of viral latency genes and these genes alter host transcription programs. To globally measure the transcriptome changes during EBV transformation, primary human resting B lymphocytes (RBLs) were infected with B95.8 EBV for 0, 2, 4, 7, 14, 21, and 28 days, and polyA plus RNAs were analyzed by RNA-seq. ANOVA analyses found 3669 protein coding genes that were differentially expressed (FDR<0.01). 94% of LCL genes that are essential for LCL growth and survival were differentially expressed. Pathway analyses identified significant enrichment of pathways involved in cell proliferation, DNA repair, metabolism, and anti-viral responses. RNA-seq also identified lncRNAs differentially expressed during EBV infection. CRISPRi and CRISPRa found that CYTOR and NORAD lncRNAs were important for LCL growth. During EBV infection, type III EBV latency genes were expressed rapidly after infection. Immediately after LCL establishment, EBV lytic genes were also expressed in LCLs and ~4% of the LCLs express gp350. ChIP-seq and POLR2A ChIA-PET data linked EBV enhancers to 90% of EBV-regulated genes. Many genes were linked to enhancers occupied by multiple EBNAs or NF-κB subunits. Incorporating these assays, we generated a comprehensive EBV regulome in LCLs. [ABSTRACT FROM AUTHOR]

Gene regulation by trans-acting small RNAs (sRNAs) has considerable advantages over other gene regulation strategies. However, synthetic sRNAs mainly take natural sRNAs (MicC or SgrS) as backbones and comprise three functional elements folding into two or more stem-loop structures: an mRNA base pairing region, an Hfq-binding structure, and a rho-independent terminator. Due to limited numbers of natural sRNAs and complicated backbone structures, synthetic sRNAs suffer from low activity programmability and poor structural modularity. Moreover, natural sRNA backbone sequences may increase the possibility of unwanted recombination. Here, we present a bottom-up approach for creating structure defined single-stem loop small non-coding RNAs (ssl-sRNAs), which contain a standardized scaffold of a 7 bp-stem-4 nt-loop-polyU-tail and a 24 nt basing pairing region covering the first eight codons. Particularly, ssl-sRNA requires no independent Hfq-binding structure, as the polyU tail fulfills the roles of binding Hfq. A thermodynamic-based scoring model and a web server sslRNAD (http://www.kangzlab.cn/) were developed for automated design of ssl-sRNAs with well-defined structures and programmable activities. ssl-sRNAs displayed weak polar effects when regulating polycistronic mRNAs. The sslsRNA designed by sslRNAD showed regulatory activities in both Escherichia coli and Bacillus subtilis. A streamlined workflow was developed for the construction of customized ssl-sRNA and ssl-sRNA libraries. As examples, the E. coli cell morphology was easily modified and new target genes of ergothioneine biosynthesis were quickly identified with ssl-sRNAs. ssl-sRNA and its designer sslRNAD enable researchers to rapidly design sRNAs for knocking down target genes. [ABSTRACT FROM AUTHOR]

In this paper, a variable-coefficient time fractional heat-like and wave-like equation with initial and boundary conditions is solved by the use of variable separation method and the properties of Mittag-Leffler function. As a result, exact solutions are obtained, from which some known special solutions are recovered. It is shown that the variable separation method can also be used to solve some others time fractional heat-like and wave-like equation in science and engineering. [ABSTRACT FROM AUTHOR]

In this paper, a direct algorithm of the exp-function method is proposed for exactly solving non-linear evolution equations. To illustrate the validity and advantages of the algorithm, the Korteweg-de Vries and Jimho-Miwa equations are considered. As a result, exact solutions are obtained. It is shown that the exp-function method with the direct algorithm provides a simpler but effective mathematical tool for constructing exact solutions of non-linear evolution equations in fluids. [ABSTRACT FROM AUTHOR]

The article looks at a study which developed a kinetic diffusion–controlled method and synthesized intermetallic Pt2Mo nanocrystals with twin boundaries on mesoporous carbon. Topics discussed include formation of small-sized twinned intermetallic nanocrystals is associated with the strong Mo-C interaction; in situ aberration-corrected transmission electron microscope (TEM) at high temperature and present strategy paves the way for tailoring the interface structure of high–melting point.

The article discusses the poor understanding of the complex multistep process taken by nanocarriers for the delivery process limits the delivery efficiencies and further hinders the translation of systems into medicine. Topics include anocarriers that have endowed with tubular shape, short length, and low rigidity outperformed the other types; and the data, simulations, and models have helpful to unseal and predict nanocarrier perform in multistep delivery for anticancer therapy.

Fermented porridge is usually made though the process of natural fermentation of grains with higher starch. Organic acids are the main flavor compounds in fermented porridge. However, the understanding of the correlation between the microbial communities and organic acids is limited in the fermentation process of traditional Chinese sour porridge (CSP). In this study, the dynamic of bacterial community and organic acids was investigated and the correlation between bacterial communities and organic acids was revealed during the fermentation process of CSP. The results showed that the richness and diversity of the bacterial communities at initial state (0h) were significantly higher than other period and the concentration of 9 organic acids (oxalic acid, tartaric acid, malic acid, lactic acid, acetic acid, citric acid, and succinic acid) increased with the fermentation process. Pediococcus and Lactobacillus were the dominative genus in the fermentation process of CSP and the fermentation process of CSP can be divided into three stages, with 0 h as the initial stage, 6-18 h as the second group and 24-36 h as the third based on PCA and cluster analysis. Moreover, 20 bacteria genus had significant correlation with (|ρ|>0.7) organic acids based on O2PLS analysis and The Lactobacillus and Acinetobacter were positively correlated with seven (succinic acid, tartaric acid, lactic acid, oxalic acid, acetic acid, malic acid and succinic acid) and four (tartaric acid, acetic acid, lactic acid, and a-ketoglutarate) organic acids respectively. Clostridium, Paenibacillus, Cupriavidus, Shinella, Streptococcus, Flavobacterium, Enterobacteriaceae, Cyanobacteria, Pseudomonas, Ralstonia, Microbacterium, Bradyrhizobiaceae, Escherichia-Shigella, Acidovorax, Leuconostoc, Alphaproteobacteria, Pediococcus, and Mitochondria were negatively correlated with organic acids in the fermentation process of CSP. [ABSTRACT FROM AUTHOR]

Early diagnosis of nasopharyngeal carcinoma (NPC) is difficult because of a lack of specific symptoms. Many patients have advanced disease at diagnosis, and these patients respond poorly to treatment. New treatments are therefore needed to improve the outcome of NPC. To better understand the molecular pathogenesis of NPC, here we used an NPC cell line in a genome-wide CRISPR-based knockout screen to identify the cellular factors and pathways essential for NPC (i.e. dependence factors). This screen identified the Moz, Ybf2/Sas3, Sas2, Tip60 histone acetyl transferase complex, NF-κB signaling, purine synthesis, and linear ubiquitination pathways; and MDM2 proto-oncogene as NPC dependence factors/pathways. Using gene knock out, complementary DNA rescue, and inhibitor assays, we found that perturbation of these pathways greatly reduces the growth of NPC cell lines but does not affect growth of SV40-immortalized normal nasopharyngeal epithelial cells. These results suggest that targeting these pathways/proteins may hold promise for achieving better treatment of patients with NPC. [ABSTRACT FROM AUTHOR]

The serine/threonine kinase Unc-51-like kinase-1 (Ulk1) is thought to be essential for induction of autophagy, an intracellular bulk degradation process that is activated by various stresses. Although several proteins have been suggested as Ulk1 substrates during autophagic process, it still remains largely unknown about Ulk1's physiological substrates. Here, by performing in vitro and in vivo phosphorylation assay, we report that the co-chaperone cell division cycle protein 37 (Cdc37) is a Ulk1 substrate. Ulk1-mediated phosphorylation of Ser-339 in Cdc37 compromised the recruitment of client kinases to a complex comprising Cdc37 and heat shock protein 90 (Hsp90) but only modestly affected Cdc37 binding to Hsp90. Because the recruitment of protein kinase clients to the Hsp90 complex is essential for their stability and functions, Ser-339 phosphorylation of Cdc37 disrupts its ability as a co-chaperone to coordinate Hsp90. Hsp90 inhibitors are cancer chemotherapeutic agents by inducing depletion of clients, many of which are oncogenes. Upon treatment with an Hsp90 inhibitor in cancer cells, Ulk1 promoted the degradation of Hsp90-Cdc37 client kinases, resulting in increased cellular sensitivity to Hsp90 inhibitors. Thus, our study provides evidence for an anti-proliferative role of Ulk1 in response to Hsp90 inhibition in cancer cells. [ABSTRACT FROM AUTHOR]

Different experimental models of hepatocellular carcinoma (HCC) have been used to investigate the biological mechanisms of hepatocarcinogenesis and its progression. However, previous studies have highlighted the difficulty of distinguishing between the tumor cells and stroma in experimental models of HCC. Therefore the aim of the present study was to establish a red-green dual-color fluorescence tracing orthotopic transplantation model of HCC, and investigate its practical values. Stable high red fluorescent protein (RFP)-expressing HepG2 human hepatoma cells and Hepa1-6 mice hepatoma cells were injected into the right liver lobe of green fluorescent protein-expressing nude mice. The growth and metastasis of the tumors were visualized using a whole-body in vivo fluorescence imaging system in real time. HCC tissues were extracted from tumor-bearing mice, and cut into 5-µm serial frozen slices. The organizational structure of the transplanted tumors was observed under a microscope. A dual-color fluorescence tracing orthotopic transplantation tumor model of HCC was successfully established with a success rate of 100%. The growth and metastasis of the tumors were visualized at each stage of development in the tumor-bearing mice. Tumor cells with red fluorescence and host cells with green fluorescence were identified to merge in the reconstruction region of tumor tissue. The invasion, migration, and cell fusion between tumor and host cells was observed clearly. The dual-color fluorescence tracing orthotopic transplantation model of HCC was determined to be a stable and reliable method for tracking tumor progression. Mutual interactions between hepatoma cells and host tissues may be observed directly using this model, further elucidating the development of the tumor microenvironment. [ABSTRACT FROM AUTHOR]

C5H7N3O4, monoclinic, P1̄ (no. 2), a = 5.1166(12) Å, b = 8.711(2) Å, c = 8.744(2) Å, α = 103.169(7)°, β = 90.454(6)°, γ = 96.121(7)°, V = 377.08(16) Å3, Z = 2, Rgt(F) = 0.0619, wRref(F2) = 0.1241, T = 173(2) K. [ABSTRACT FROM AUTHOR]

C12H5Cl2N5O7, triclinic, P1̄ (no. 2), a = 8.2425(8) Å, b = 8.2814(7) Å, c = 12.4713(12) Å, α = 93.370(4)°, β = 101.590(4)°, γ = 118.222(3)°, V = 723.03(12) Å3, Z = 2, Rgt(F) = 0.0516, wRref(F2) = 0.0858, T = 173(2) K. [ABSTRACT FROM AUTHOR]